Faculty of Pharmacyhttp://khartoumspace.uofk.edu/123456789/269612020-06-07T07:18:52Z2020-06-07T07:18:52ZAntiviral Activity of Acacia nilotica Against Influenza AVirusGassab, Mona Timan Idrisshttp://khartoumspace.uofk.edu/123456789/271222019-09-15T07:36:54ZBackground: Human influenza is a seasonal disease associated with significant morbidity and mortality.The emergence of drug-resistant virus strains highlights the urgent need to develop novel antiviral drugs with alternative modes of action. Anti-flu Traditional Chinese Medicine (TCM) has played a significant role in fighting the virus pandemic. In Sudan, Acacia nilotica is a commonly used ingredient in many therapeutic remedies, either alone or in conjunction with other natural substances. Evidence suggests that Acacia nilotica is associated with a variety of pharmacological activities.
Object of this studywas to investigate the influence of Acacia niloticamethanolic extract on the infectivity of influenza viruses in Madin–Darby Canine Kidney (MDCK) cells, and its possible in vitro mechanism(s) of action.
Methodology: The fruit husk of Acacia niloticawas collected from Sudan and extracted with70% methanol. The crude extract was screened for its cytotoxicity against MDCK cell line by crystal violet assay and Water Soluble Tetrazolium-1 (WST-1) assay. Antiviral properties of Acacia niloticaextract was determined by cytopathic inhibition assay, virus yield reduction assay (plaque assay), nuclear export inhibitor and western blotting assay. To assess the possible mechanism of action, time of addition assay was performed targeting different sites in virus life cycle.
Result:Acacia nilotica has been found to have anti-influenza-virus activity, and both pre-incubation of virus prior to infection and post-exposure of infected cells with Acacia nilotica extracts significantly inhibited virus yields. Influenza-virus-induced hemagglutination of chicken red blood cells was inhibited by Acacianilotica extract, suggesting that Acacia nilotica can inhibit influenza A virus infection by interacting with the viral hemagglutinin. Furthermore, Acacianilotica extract significantly affects nuclear transport of viral nucleoprotein (NP).
Conclusion: The antiviral activity of Acacia niloticaextracts indicates that a component or components of these extracts possess anti-influenza virus properties. To the best of our knowledge, this study revealed for the first time that Acacia nilotica extract can inhibit both viral attachment and replication and offers new insights into its underlying mechanisms of antiviral action.
Antiviral Activity of Acacia nilotica Against Influenza AVirus
In vitro Glycogen Phosphorylase Inhibition Coupled with LC/MS Profiling and Antioxidant Activity of Nauclea latifolia Smith FruitsAbd Algaffar, Shereen Omer Elfaroukhttp://khartoumspace.uofk.edu/123456789/270352019-07-08T09:51:43ZBackground: The long-term effects of the current clinically used anti-diabetic
drugs are still unsatisfactory with various adverse effects which limit their usage.
Considering its central role in glycogen metabolism, glycogen phosphorylase
(GP); an allosteric protein with at least six different ligand binding sites that
modulate its enzymatic activity; has recently emerged as an important therapeutic
target to discover potent antidiabetic drugs for type 2 diabetes (T2D). Meanwhile,
recent evidence indicates the intricate association of the inflammatory damage
characterizes diabetes and the pathophysiological events mediated through
pancreatic islets reactive oxygen species (ROS) in the loss of β-cell function.
Traditional medicine in Sudan have been used for centuries to treat diseases
including diabetes, hence there are already vast literatures available for data
mining and among them Nauclea latifolia fruit (Karmadoda) standing very
prominently. Therefore, during the last few years, considerable attention has been
given to scientifically validate the bioactivity of some antidiabetic traditional
plants.
The aim of this study is to validate the traditional use of N. latifolia fruit as
antidiabetic agent and identify its chemical components, molecular target, and
mechanism of action in comparison with the intact fruits.
Methods: The plant including its fruits was taxonomically identified as N.latifolia
Smith and herbarium specimens were deposited. Both the intact and processed
fruits were extracted with 70% ethanol, concentrated, and fractionated with
petroleum ether, chloroform and ethyl acetate respectively. The crude extract and
all its respective fractions were screened for (GP) inhibition using GPa. Caffeine
and quercetin were used as standards. The Antioxidant activity was performed
according to standard methods using DPPH (2,2 Diphenylpicrylhydrazyl) and
ABTS (2,2’- Azinobis( 3-benzothiazoline- 6 sulfonic acid). Scavenging capacity
of the compounds was compared with that of Ascorbic acid and Trolox.
The quantification of the flavonoids contents was executed according to
Aluminum chloride colorimetric assay method and total flavonoid content was
expressed as quercetin equivalent per 100g dry weight of sample. The total
phenolic content was determined using Folin-Ciocalteu reagent and the results
were expressed as gallic acid equivalents per 100g dry plant matter. LC-PDAESIMS analysis were performed on apparatus with an electrospray source assisted
by the DataAnalysis® software. The online SciFinder Nauclea genus database
“Nauclea DB” was used to predict from all NPs of Nauclea genus. SpectrusX
processor, C+HNMR Predictors and DB software (ACD/Labs, 2014 Release) were
used to predict the chemical shifts of these natural products. Molecular Operating
Environment (MOE. v. 2008.10, Chemical Computing Group, Canada) and
Discovery Studio Visualizer Softwares were used for the visualization of the
docking results and generating the images.
Results: Systematic screening of N.latifolia Smith intact fruit and its traditionally
processed fruit (Karmadoda) revealed a remarkable GP inhibition for the intact
fruit (95.57%) in comparison with the processed fruit. The DPPH and ABTS
radical scavenging activity revealed that the intact fruit ethyl acetate fraction
exhibited the most prominent antioxidant activity (89.50±.0..0%DPPH RSA±
SD) which far exceeding its respective processed counterpart (83.57±0.02 %DPPH
RSA± SD). Statistical analysis indicated that processing of fruits had a direct
negative significant impact (P< 0.05) on both the antidiabetic and antioxidant
activities. Quantification of both the total phenolics and total flavonoid contents
revealed higher polyphenolic contents in the intact fruits compared to processed
ones.
Planar chromatographic analysis and hyphenated LC/MS techniques resulted in
the identification of five β-carboline alkaloids, a flavonoid glycoside and
chlorogenic acid in the intact fruit, while the processed fruit exhibited different
chemical profiles characterised by the presence of only three β-carboline alkaloids
besides the flavonoid glycoside in addition to a bis (2-ethylhexyl) phthalate which
is considered as an artefact.
Eventually, among the allosteric molecular targets analysed computationally, the
catalytic site proved to be the most appropriate to accommodate both the phenolic
and alkaloids identified in the fruit as ligands with the flavonoids luteolin 3'-
methyl ether 7-glucuronide-4'-rhamnoside and chlorogenic acid scoring -28.39 and
-21.42 Kcal/mol, respectively, among the phenolic compounds, as well as the 10-
hydroxy strictosamide and strictosidine scoring - 28.40 and -23.07 Kcal /mol,
respectively.
Conclusions: The intact fruit is superior in terms of its antidiabetic and
antioxidant capacity than the processed fruit. The catalytic site has been identified
as the main N. latifolia molecular target for GP antidiabetic activity
In vitro Glycogen Phosphorylase Inhibition Coupled with LC/MS Profiling and Antioxidant Activity of Nauclea latifolia Smith Fruits
Activity- guided Isolation and Structure Determination of Antioxidant and Antidiabetic Compounds from Bauhinia rufescence L.Osman, Wadah Jamal Ahmedhttp://khartoumspace.uofk.edu/123456789/270272019-06-27T07:54:04ZBackground:
Diabetes mellitus is a group of disorders of multiple etiologies resulting from a defect in
insulin secretion, insulin action, or both. Diabetes remain one of the world’s major health
problems and many repots have been supported the role of oxidative stress in the
pathogenesis of both type 1 and type 2 diabetes.
This study aimed to evaluate the antidiabetic and antioxidant activity of Bauhinia
rufescens a plant used in Sudanese folkloric medicine for treatment of diabetes. It was also
aimed to identify isolates and characterize the bioactive antidiabetic and antioxidant
compounds using bioactivity guided fractionation followed by High Performance Thin Layer
Chromatography (HPTLC) autobiography, Liquid-Mass spectroscopy analysis and nuclear
magnetic resonance (NMR).
Method:
The powdered plant leaves were exclusively extracted using petroleum ether, chloroform and
methanol, successively.
These extracts were tested in streptozotocin-induced diabetic Wistar rats for antidiabetic as
well as DPPH antioxidant potential. Standard Propyl Gallate was used as (+ve) control and
the difference in results was considered significant when P < 0.05. The best active extract
was fractionated with chloroform, ethyl acetate and butanol along with the remaining
aqueous fraction. The fractions are tested for antioxidant and antidiabetic activity using
HPTLC autobiography (α amylase and α glucosidase).
Further, bioactive compounds were isolated by Preparative Thin Layer Chromatography and
identified using LC/MS and NMR spectroscopy.X
Results:
The methanol extract significantly decreased the blood glucose level in normal fasted rats by
27 %; two hours following administration and decreased the blood glucose level in
streptozotocin-treated rats by 48.9%; two hours following oral administration, while
chloroform and pet. Ether extracts not significantly decreases the glucose level.Significantly
the methanol extract also showed strong antioxidant activity (89%) compared with other
extracts; thereforeit was further subjected for fractionation by using chloroform, ethyl acetate,
n-butanol and water sequentially, and then tested for their antioxidant activity; where the
ethyl acetate fraction was showed good activity compared with other fractions.
Bioactivity guided fractionation of Bauhinia rufescensled to isolation and characterization
of two compounds with potent antidiabetic and antioxidant activity namely; Quercetin-3-Orutinoside and Kaempferol -3-O-rutinoside.
These compounds were isolated by using preparative thin layer c chromatographic
techniques (PTLC) and their structures were identified by 1D NMR spectrum (C13and 1HNMR), 2D NMR (COSY, HSQC, HMBC Correlation) and confirmed bycomparing with
reported data.
LC/MS analysis led to identified more than thirty compounds; which were subjected to in
silico assessment against human α-glucosidase and α-amylase enzymes. Insilico assessment
showed that more than 14 compounds showed potent activity compared with acarbose
(standard anti-diabetic drug).
High performance thin layer chromatography (HPTLC) for the first time of the active
fractionof the plant revealed the presence of different spots/peaks in different Rf values,
corresponding to different compounds.XI
Conclusion:
This study validated the traditional uses of the evaluated plant in treating diabetes; beside that
it was found a good antioxidant. The compounds which were isolated and identified may
provide a new lead for more potent analogues for drug discovery. The anti-diabetic potential
and antioxidant activity reported in present investigation is reported for the first time on the
aerial plant. Further the isolated and characterized compounds from this study are also being
reported for the first time.
Toxicological studies may further be planned to develop it in the form of new drug for
management of diabetes, however its use is available as per traditional claim.
Activity- guided Isolation and Structure Determination of Antioxidant and Antidiabetic Compounds from Bauhinia rufescence L.
Detection of Multi Drug Resistant TB Strains Using Conventional And Molecular TechniquesIsmat, Gusai Hussein Abdel Samadhttp://khartoumspace.uofk.edu/123456789/267222019-01-03T13:07:25ZBackground: - Multidrug-resistance against the first line drugs for treatment of tuberculosis arises when the patients fail to complete the prescribed doses, and when the concentration of the drugs inside their bodies is not enough to kill all of the causative bacteria .This may be attributed to stoppage of treatment by patients when they feel better or they are unable to pay for the whole regimen that takes months. The infectivity of MDRT-TB strains is not affected and they remain to be fully virulent as non - resistant strains. The study aimed to detect the multi- drug resistant TB strains using conventional and molecular techniques and to detect the mutations that may be found in an effort to decrease the time of diagnosis and to have more accurate results that lead to fast and effective treatment.
Methods:- The study was done on patients with pulmonary tuberculosis at Abu-Anga Teaching Hospital , Omdurman province during the year 2005. Samples of sputum from in-patients suffering from pulmonary tuberculosis were collected. These samples were pretreated with 4%NaOH and cultivated on L.J. media. Conventional methods (68Co labile catalase test , Nitrates reduction test , growth on PNB medium were used to identify the isolates . Collection , transport, pretreatment of samples , inoculation, incubation, reading and reporting of growth were done according to the WHO instructions, (1998). Susceptibility to rifampicin and isoniazid (proportion method) was performed for the previously obtained isolates.DNA from 20 resistant isolates was extracted . Molecular techniques such as PCR, cloning and sequencing were performed to detect the KatG gene responsible for isoniazid resistance.
Results :- During the year 2005 , 6736 patients visited the out-patient clinic of Abu-Anga Hospital, 4699 were males and 2037 females. Only 431 patients (322 males, 109 females) were admitted to the hospital. Three hundred and five isolates were obtained, identified as Mycobacterium complex. The sensitivity test for the 305 isolates showed that 238(92.8%) isolates were sensitive to both rifampicin and isoniazid. Nineteen (6.2%) isolates were resistant to both rifampicin and isoniazid .Two isolates were resistant to isoniazid only and one isolate was resistant to rifampicin only. Multidrug resistance was found in 6.2%.of the isolates. Mutation of type S315T was observed in 20 isolates.
Conclusion:- The prevalence of MDR-TB was found to be 6.2 % among clinical isolates at Abu-anga Hospital. The results were obtained in short time when using molecular techniques rather than conventional ones which take time . This provides quick results and fast treatment for MDR-TB patients.
Detection of Multi Drug Resistant TB Strains Using Conventional And Molecular Techniques